San Pedro”) and Their Relevance to Shamanic Practice

Journal of Ethnopharmacology 131 (2010) 356–362

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Journal of Ethnopharmacology

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New mescaline concentrations from 14 taxa/cultivars of Echinopsis spp. (Cactaceae) (“San Pedro”) and their relevance to shamanic practice

Olabode Ogunbodede a, Douglas McCombs a, Keeper Trout b, Paul Daley c, Martin Terry a,b,∗ a Sul Ross State University, Department of Biology, Alpine, TX 79832, USA b Cactus Conservation Institute, 909 E. Sul Ross Ave., Alpine, TX 79830, USA c California Paciﬁc Medical Center, Addiction Pharmacology Research Laboratory, P.O. Box 7999, San Francisco, CA 94120, USA article info abstract

Article history: Aim of the study: The aim of the present study is to determine in a procedurally uniform manner the Received 17 April 2010 mescaline concentrations in stem tissue of 14 taxa/cultivars of the subgenus Trichocereus of the genus Received in revised form 29 May 2010 Echinopsis (Cactaceae) and to evaluate the relationship (if any) between mescaline concentration and Accepted 6 July 2010 actual shamanic use of these plants. Available online 14 July 2010 Materials and methods: Columnar cacti of the genus Echinopsis, some of which are used for diagnostic and therapeutic purposes by South American shamans in traditional medicine, were selected for analysis Keywords: because they were vegetative clones of plants of documented geographic origin and/or because they were Traditional medicine Shamanism known to be used by practitioners of shamanism. Mescaline content of the cortical stem chlorenchyma Echinopsis of each cactus was determined by Soxhlet extraction with methanol, followed by acid–base extraction Trichocereus with water and dichloromethane, and high-pressure liquid chromatography (HPLC). San Pedro Results: By virtue of the consistent analytical procedures used, comparable alkaloid concentrations were Analytical chemistry obtained that facilitated the ranking of the various selected species and cultivars of Echinopsis, all of which Mescaline exhibited positive mescaline contents. The range of mescaline concentrations across the 14 taxa/cultivars spanned two orders of magnitude, from 0.053% to 4.7% by dry weight. Conclusions: The mescaline concentrations reported here largely support the hypothesis that plants with the highest mescaline concentrations – particularly E. pachanoi from Peru – are most associated with documented shamanic use. © 2010 Elsevier Ireland Ltd. All rights reserved.

1. Introduction

The portion of the genus Echinopsis (Cactaceae) that was for- Abbreviations: AUFS, absorbance units full scale; C, centigrade; ca., circa (approx- merly known as Trichocereus (Friedrich, 1974; Rowley, 1974) imately); cf., confer (compare); cm3, cubic centimeter; CAS, Chemical Abstracts consists of largely columnar cacti native to the Andean slopes Service; Cl, chloro; CSA 1, Schedule 1 in the US Controlled Substances Act; cv., cultivar; E., Echinopsis; Dept., Department; et al., et alii (and others); FR, a col- of Ecuador, Peru, northern Chile, and Bolivia. Several species lection number of Friedrich Ritter; g, gram; GC–MS, gas chromatography–mass of Echinopsis are used for diagnostic and therapeutic purposes spectroscopy; GE, substance listed in German Controlled Substances Act (BtMG); by indigenous practitioners. Such use is evidently based on HPLC, high-performance liquid chromatography; IC, substance under international their psychoactive properties due to their mescaline content control (United Nations); ID, internal diameter; in litt., in litteris (in a note or email); IOS, International Organization for Succulent Plant Studies; KK, Karel Knize; (Dobkin de Ríos, 1972). The species most widely and frequently L, liter; M, mescaline; m, meter; mg, milligram; mL, milliliter; MSD, mass-selective used in this manner, viz., Echinopsis pachanoi (Britton and Rose) detection; MW, molecular weight; m/z, mass-to-charge ratio; ␮L, microliter; ␮g, Friedrich and Rowley (E. pachanoi), Echinopsis peruviana (Britton microgram; N, normal; N/A, not available; nm, nanometer; NMCR, New Mexico Cac- and Rose) Friedrich and Rowley (E. peruviana) and Echinop- tus Research; OST, a collection number of Carlos Ostolaza; PCH, a collection number sis lageniformis (Foerster) Friedrich and Rowley (E. lageniformis), of Paul C. Hutchison; PIH, PIHKAL (Shulgin and Shulgin1991); pKa, ionization con- stant; Prov., Province; QI, quality index of spectra; Rd., road; REF, reference; RI, gas are often referred to in Spanish as San Pedro, alluding to St. chromatographic Kovats retention index; SE 30, a standard GC column; s.l., sensu Peter’s role as the gatekeeper to heaven (Schultes and Hofmann, lato; sp., species; s.s., sensu stricto; ssp., subspecies; T., Trichocereus; TSQ, a MS data 1980). system; UV, ultraviolet; viz., videlicet (namely); w/w, weight per weight (expression The single most commonly utilized species, E. pachanoi, has of concentration). ∗ been cultivated for such a long time that it is difﬁcult to determine Corresponding author at: Sul Ross State University, Department of Biology, Alpine, TX 79832, USA. Tel.: +1 432 837 8113; fax: +1 432 837 8682. its geographic origin and natural habitat (Britton and Rose, 1920; E-mail addresses: [email protected], [email protected] (M. Terry). Yetman, 2007).

Earlier analytical and pharmaceutical reports identified spec- Due to the difficulty of obtaining reliably documented specimens chemically assayed as Opuntia cylindrica (Cruz Sánchez, imens, we decided at the outset to analyze only one individual 1948; Marini-Bettòlo and Coch Frugoni, 1958; Turner and Heyman, to represent each taxon/cultivar. Thus, the study design expressly 1960). Friedberg (1959) was the first investigator to report that ignores the (probably considerable) variation in mescaline con- the ritual drug plant popularly known as San Pedro corresponds tent among individuals within a given taxon. Temporal variation to that described by Britton and Rose (1920) as Trichocereus in mescaline content that may be associated with age/size of pachanoi (=Echinopsis pachanoi). Poisson (1960) was the first to the plant, season of the year or time of day, was also not con- isolate mescaline from a San Pedro validly identified as Tri- trolled. Living voucher specimens of cacti used in this study chocereus (Echinopsis) pachanoi, using Friedberg’s (1959) voucher are being maintained in our greenhouse. Photographs of these material. specimens can be found at www.cactusconservation.org/botany/ While there seems to be universal consensus among scien- Ogunbodede.html. tists that mescaline is the active principle in Echinopsis species A fresh sample of chlorenchyma from the green outer cortex underlying their use, the mescaline content of these cacti has long of the stem of each Echinopsis species to be assayed was weighed, been a subject of controversy. Different authors employing various sliced and cut into cubes ca. 1 cm3, desiccated for ca. 30 h at room methods and instrumentation have published markedly different temperature, and weighed in the dried state. The dried material mescaline contents for these plants (Table 1). was finely ground with mortar and pestle. Two (2.0) grams of the Part of the variation among these published results for the sub- dry, ground chlorenchyma tissue was extracted with methanol in genus Trichocereus of the genus Echinopsis may be attributable to a Soxhlet apparatus for 8 h at 40 ◦C. The extract was evaporated to real genetic differences in the regulation of mescaline biosynthe- dryness at 40 ◦C in a rotary evaporator, redissolved in 150 mL of sis among the various species/cultivars sampled, as well as among distilled water, and acidified with concentrated hydrochloric acid populations within a given species/cultivar and individuals within to pH 3.0. The acidified aqueous extract was defatted twice with a given population. Environmental factors, including variation in 50 mL of dichloromethane. The aqueous layer was alkalinized to temperature and rainfall (which correlate with differences in alti- pH 12.0 with 5 N sodium hydroxide (the pKa of the protonated tude), as well as edaphic conditions, could also be expected to amino group of mescaline being 9.5), and extracted twice with contribute to geographic and/or temporal variation in mescaline 50 mL dichloromethane. The aqueous layer was discarded, and the content. But it was suspected that a significant part of the reported dichloromethane evaporated to dryness. The extract was redis- variation in mescaline content of E. pachanoi and related taxa might solved in 10.0 mL of methanol, filtered through a 0.2-␮m micropore be attributable to interlaboratory differences in technique. This filter to remove particulates, and stored in a glass vial at 4 ◦C, pend- latter source of variation amounts to noise that may seriously con- ing high-pressure liquid chromatography (HPLC). found previously reported results, precluding a valid comparison among them. It is also noteworthy that much of the published 2.2. Analytical instrumentation and methodology research in this area dates to the 1960s and 1970s, when differences among laboratories were likely greater than at present. There The HPLC instrumentation consisted of a Beckman 322 is therefore the need for confirmatory studies to evaluate these gradient liquid chromatograph fitted with a Beckman 110A earlier results. solvent-metering pump, Beckman 421 controller, a Spectra 100 The primary objective of this project was thus to employ mod- variable-wavelength detector (Spectra Physics), and a Kipp and ern, uniform analytical methods in a single laboratory, to determine Zonen BE 8 multirange recorder. Alkaloid separation was carried the mescaline contents of E. pachanoi and related taxa, includ- out isocratically with a Phenomenex Luna 3-␮m 250 mm × 4.6 mm ing some cultivars. All analyses were conducted by the same ID reverse phase C-18 column at 25 ◦C. The mobile phase con- investigator (Ogunbodede) using the same procedures, the same sisted of 10.8% acetonitrile, 89.2% water, acidified with 0.10% experimental conditions, and the same equipment. A secondary trifluoroacetic acid, at a flow rate of 0.5 mL per min. The detec- objective was to examine the relationship, if any, between mesca- tor wavelength was set at a known UV absorbance maximum of line concentration in Echinopsis taxa/cultivars and any documented 205 nm (as per Helmlin and Brenneisen, 1992). use of those taxa/cultivars by indigenous shamanic practitioners. The HPLC fractions were injected into an Agilent 6890 gas The hypothesis was that the cacti with the highest mescaline tis- chromatograph (GC) with a 5972 mass-selective detector (MSD) sue concentrations would be more likely to be used in shamanic operating in 70 eV electron ionization mode, using Agilent Chem- practice and that such use would be reflected in the literature. station (v. C.00.00) data acquision software. Mass spectra were obtained with the Chemstation analysis utilities, and compared with both standards and library spectra (using MSSearch 2.0, NIST, 2. Materials and methods 1998). Authentic mescaline hydrochloride (Grace Davison, purchased 2.1. Plant selection and extraction by M. Terry, DEA Researcher Registration No. RT 0269591) was used as the mescaline reference standard. The concentrations of the In addition to the taxonomic uncertainty that surrounds the standard solutions and the corresponding mean HPLC peak heights genus Echinopsis, there is ethnobotanical uncertainty as to how the were plotted in a standard curve and correlated using a Labworks various species and cultivars are used by practitioners of differ- spreadsheet. The range of mescaline concentrations covered by the ent cultures over the large geographic area to which the genus is standard solutions was 4.59–73.3 ␮g/mL. A dataset was considered endemic. Accordingly, the criteria for selecting plants for analy- acceptable if the correlation coefficient was ≥0.999. sis were that (a) the plants had a reasonable amount of credible botanical documentation as to their collection data, with emphasis on geographic origin, and/or (b) the plants exhibited documented 3. Results use for shamanic therapeutic purposes. The exception was the single case of E. pachanoi cv. Juul’s Giant. This plant is documented as All specimens analyzed contained detectable levels of mesca- a cultivar in the United States, but the geographic region of Peru to line (Table 2). The mescaline HPLC peaks of all samples analyzed which it is native is not known. It bears close resemblance to plants exhibited a consistent retention time of 14 min. Mescaline HPLC encountered around Arequipa (Trout, 2005). peak confirmation was first demonstrated by co-elution with the 358 O. Ogunbodede et al. / Journal of Ethnopharmacology 131 (2010) 356–362

Table 1 Reported concentrations (in %) of mescaline in Echinopsis cacti with afﬁnities to E. pachanoi and E. peruviana. CP: chlorophyllaceous parenchyma only. WP: whole plant.

Species % Form/part Locale or source Reference

E. lageniformis >0.25 Dry/WP Horticulture, European Agurell (1969b) 0.56 Dry/CP La Paz, Bolivia Serrano (2008)

E. cuzcoensis 0.0 Dry/CP Cotaruse, Arequipa, Peru Serrano (2008) 0.0 Dry/CP Huaytampo, Cuzco, Peru Serrano (2008) 0.0 Dry/CP Huacarpay, Cuzco, Peru Serrano (2008) 0.0 Dry/CP Capacmarca, Cuzco, Peru Serrano (2008) 0.005–0.05 Fresh/WP Horticulture, Germany Agurell et al. (1971)

E. pachanoi 5 Dry/CP Cultivated, Lima, Peru Cruz Sánchez (1948) 4.5 Dry/CP Cultivated: Lima, Peru Gonzalez Huerta (1960) 2.06 Dry/WP Horticulture, Italy Gennaro et al. (1996) 0.109–2.375 Dry/WP Horticulture, Switzerland Helmlin and Brenneisen (1992) 1.2 Dry/WP Huancabamba, Peru Poisson (1960) 0.9 Dry/WP Peruvian drug material Turner and Heyman (1960) 0.331 Dry/WP Horticulture, California Crosby and McLaughlin (1973) >0.025 Fresh/WP Horticulture, European Agurell (1969b) 0.04–0.067 Fresh/WP Horticulture, European Agurell (1969a) 0.067–0.079 Fresh/WP Horticulture, European Bruhn and Lundström (1976) 0.15–0.155 Dry/WP Horticulture, California Pummangura et al. (1982) 0.78 Dry/CP Chiclayo, Peru Reyna Pinedo and Flores Garcés (2001) 1.4 Dry/CP Barranca, Peru Reyna Pinedo and Flores Garcés (2001) 0.00 Dry/CP El Alisal, San Marcos, Cajamarca, Peru Cjuno et al. (2009) 0.00 Dry/CP Cataratas, Otuzco, La Libertad, Peru Cjuno et al. (2009) 0.23 Dry/CP Moyán, San Vincente, Lambayeque, Peru Cjuno et al. (2009) 0.28 Dry/CP Puykate, Ferrenafe,˜ Lambayeque, Peru Cjuno et al. (2009) 0.38 Dry/CP Yanasara, Sánchez Carrión, La Libertad, Peru Cjuno et al. (2009) 0.45 Dry/CP KunturWasi, San Pablo, Cajamarca, Peru Cjuno et al. (2009) 0.94 Dry/CP Tocmoche, Chota, Cajamarca, Peru Cjuno et al. (2009) 1.14 Dry/CP Laquipampa, Ferrenafe,˜ Lambayeque, Peru Cjuno et al. (2009)

E. peruviana 0.25 Dry/CP Chavin de Huantar, Huari, Ancash, Peru Cjuno et al. (2009) 0.0 Dry/WP Horticulture, European Agurell (1969b) 0.0 Dry/WP Wild harvested in Peru Djerassi et al. (1955) 0.817 Dry/WP KK242 seed from Matucana, Peru grown in California Pardanani et al. (1977)

E. puquiensis 0.28 Dry/CP Chavina,˜ Lucanas, Ayacucho, Peru Serrano (2008) and Cjuno et al. (2009) 0.13 Dry/CP Chumpi, Parincochas, Ayacucho, Peru Serrano (2008) and Cjuno et al. (2009) 0.11 Dry/CP Incuyo, Parincochas, Ayacucho, Peru Serrano (2008) and Cjuno et al. (2009) 0.50 Dry/CP Vado, Lucanas, Ayacucho, Peru Serrano (2008) and Cjuno et al. (2009)

E. santaensis 0.31 Dry/CP Mancos, Yungay, Ancash, Peru Cjuno et al. (2009)

E. schoenii 0.22 Dry/CP Cotahuasi, La Unión, Arequipa, Peru Serrano (2008) and Cjuno et al. (2009) 0.20 Dry/CP Pampacola, Castilla, Arequipa, Peru Serrano (2008) and Cjuno et al. (2009) 0.14 Dry/CP Huambo, Arequipa, Peru Serrano (2008) and Cjuno et al. (2009)

Table 2 Identity, mescaline content, collection number (if any) or geographic origin, and material originally collected (seed or cutting) of Echinopsis (Cactaceae) species/cultivars examined, in order of decreasing mescaline content.

Cactus species/cultivar Mescaline conc. (% of dry weight of cactus tissue) Collection number or origin, material originally collected

E. pachanoi 4.7 Matucana, Lima Region, Peru Cuttinga E. pachanoi cv. Juul’s Giant 1.4 Cultivar Cutting E. pachanoi (long spined) 1.2 Huancabamba, Piura Region, Peru Seed (Van Geest) E. scopulicola 0.85 FR 991: Tapecua, O’Connor Prov., Bolivia Seed (Hildegard Winter) E. pachanoi 0.82 Hutchison et al. 6212: Rio Maranon˜ above Chagual, La Libertad Dept., Peru Cutting E. pachanoi (short spined) 0.54 Huancabamba, Piura Region, Peru Seed (Van Geest) E. lageniformis (monstrose) 0.48 Cultivar Cutting E. pachanoi complex cf. T. pallarensis Ritter 0.47 FR 676: Pallar, Ancash Dept., Peru Seed (Hildegard Winter) E. pachanoi complex cf. T. riomizquensis Ritter 0.40 FR 856: Chuyllas, on the Rio Mizque, Prov. Campero, Bolivia Seed (Hildegard Winter) E. santaensis 0.32 OST 92701: Santa Valley, Ancash Dept., Peru Seed (Ostolaza) E. peruviana 0.24 KK 242: Matucana, Lima Region, Peru Cutting from Karel Knize E. lageniformis 0.18 Cultivar Cutting (Gillette) E. puquiensis 0.13 P.C. Hutchison 1256A: Nazca-Puquio Rd., across canyon from Pachan, Ayacucho Dept., Peru Cutting E. uyupampensis 0.053 Backeberg (Monaco #3487) Cutting

a Collector was an indigenous supplier of E. pachanoi to traditional Peruvian shamans’ markets, who requested anonymity. O. Ogunbodede et al. / Journal of Ethnopharmacology 131 (2010) 356–362 359

Fig. 1. GC–MS conﬁrmation of mescaline in the mescaline HPLC peak from the extract of Echinopsis pachanoi (Matucana). Note the 211 mass peak of the molecular ion, as well as the characteristic mass peaks at 194, 181 and 167. The mass peak at 223 is a common artifact (see Fig. 2). mescaline standard (Grace Davison). When the mescaline HPLC that with this instrument, in addition to the expected molecular peak obtained for a sample of extract of E. pachanoi (Matucana) mass of 211 there were apparent adduct ions detected at an m/z tissue was analyzed by gas chromatography–mass spectrometry of 223. Examination of standard runs with this system revealed (GC–MS), the mass spectrum showed the characteristic molecular that this adduct was not always present, and was not accompanied ion mass peak and fragment peaks for mescaline (Fig. 1). We noted by any shift in chromatographic behavior (retention time, slight 360 O. Ogunbodede et al. / Journal of Ethnopharmacology 131 (2010) 356–362

Fig. 2. Recent mass spectrum of mescaline by Rösner et al. (2007). Note the artifactual peak at m/z 223, which we also observed. Abbreviations: CAS, Chemical Abstracts Service; Cl, chloro; CSA 1, Schedule 1 in the US Controlled Substances Act; GE, substance listed in German Controlled Substances Act (BtMG); IC, substance under international control (United Nations); M, mescaline; MW, molecular weight; m/z, mass-to-charge ratio; PIH, PIHKAL (Shulgin and Shulgin, 1991); QI, quality index of spectra; REF, reference; RI, gas chromatographic Kovats retention index; SE 30, a standard GC column; TSQ, a popular MS data system. peak asymmetry). Similar mass spectroscopy results (Fig. 2) were stem chlorenchyma on a dry-weight basis (Table 2). The other, obtained by Rösner et al. (2007), although these data were obtained E. pachanoi (Huancabamba short spine), showed a mescaline con- with a chemical ionization instrument. We conclude that the MSD centration of 0.54% (Table 2). This difference in mescaline content did reliably identify mescaline, yet note there appears to be some between two plants of common origin – both derived from cuttings ionization chemistry taking place that is not entirely understood. of plants grown from seed collected by Dick Van Geest in the 1960s – is particularly interesting in that it shows a greater-than-twofold 4. Discussion difference in mescaline, representing the extent of individual variation among individuals produced from field-collected seed. The analytical results of the Echinopsis species and cultivars in Ontogenic variation (age and degree of maturity of the plants), Table 2 clearly indicate that the specimens showing the great- environmental variation (differences in horticultural conditions), est concentrations of mescaline in dry stem chlorenchyma tissue and temporal (seasonal and diurnal/nocturnal) variation in rates of belong to (or have affinities with) the species E. pachanoi. This alkaloid biosynthesis and degradation could also contribute to the elevated mescaline content raises interesting questions about the observed variation in the mescaline levels. variation observed in the use of this species for religious and heal- The E. peruviana cultivar examined in this study, E. peruviana KK ing purposes dating back to ancient times among various cultures 242, yielded a comparatively low value of 0.24% mescaline content of Peru and other countries of the Andes region. The markedly high on a dry-weight basis (Table 2). Curiously, an earlier E. peruviana mescaline concentration of E. pachanoi (Matucana) in the present analysis did not detect any presence of mescaline (Agurell, 1969b). study, at 4.7%, could be attributed in part to the genetic capacity The reason for this absolute difference in results could be varietal of the Matucana cultivar, but another important consideration is or procedural. There are many diverse cultivars of E. peruviana, and the analysis of only the chlorophyllaceous outer layer of the corti- anecdotal human bioassay accounts suggest widely variable mesca- cal parenchyma (i.e., chlorenchyma). This tissue has been reported line concentrations (Trout, 2005). However, Pardanani et al. (1977) to exhibit a higher mescaline concentration than the rest of the reported a mescaline recovery of 0.82% (dry weight) for seed-grown cactus stem (Janot and Bernier, 1933; Reti and Castrillón, 1951). E. peruviana KK 242, which is similar to the values for several of the Cruz Sánchez (1948), Gonzalez Huerta (1960), Serrano (2008) and other Echinopsis species/cultivars analyzed in this study, though not Cjuno et al. (2009), other workers analyzed the outer layer sepa- for E. peruviana, in the present work. =/ rately in order to overcome the difficulties posed by mucilage in Two cultivars of E. lageniformis (=Trichocereus bridgesii E. the isolation of alkaloids. Our result of 4.7% for E. pachanoi (Matu- bridgesii Salm-Dyck) were analyzed for mescaline content. E. cana) agrees very well with those of Cruz Sánchez (1948),at5%, lageniformis (monstrose) showed 0.48% mescaline content on a and Gonzalez Huerta (1960), at 4.5%. Both of those workers ana- dry-weight basis (Table 2). E. lageniformis (Gillette) showed 0.18% lyzed E. pachanoi from the Lima Region of Peru, and both analyzed mescaline content on a dry-weight basis (Table 2). By comparison, only stem chlorenchyma. While the concentration of mescaline Agurell (1969b) reported a mescaline content of about 25 mg per in chlorenchyma tends to be higher than that in the intact stem 100 g of fresh weight for intact stem tissues of E. lageniformis,an as would be used for ingestion by shamans, the selection of only amount equivalent to about 0.4% mescaline content in dry tissue. the chlorenchyma layer for analysis permits direct comparison Serrano (2008) recently reported a mescaline content of 0.56% for between species. E. lageniformis from the eastern side of La Paz, Bolivia, at an alti- Trout (2005) stated that a common error embedded in the liter- tude of 3400 m. These published figures for E. lageniformis compare ature is reference to E. peruviana as being 10 times more “potent” favorably with the figure reported for the mescaline content of E. than E. pachanoi. Our results indicate that E. pachanoi contains a lageniformis (monstrose) in Table 2. considerably higher concentration of mescaline than E. peruviana Recently Serrano (2008) published some quantitative data on (Table 2). To add support to the notion that some E. pachanoi culti- the mescaline content of Echinopsis puquiensis (Rauh and Backe- vars can be very high in mescaline content, Helmlin and Brenneisen berg) Friedrich and Rowley (E. puquiensis), harvested from different (1992) spectroscopically assayed six Swiss cultivars of E. pachanoi altitudes and locations in Peru (Table 1), and reported values which obtained from retail and private collections, and reported that one largely compare favorably with the mescaline content of 0.13% Swiss-grown E. pachanoi specimen had a mescaline concentration determined for E. puquiensis in this study (Table 2). Similarly, the 22 times greater than other Swiss-grown E. pachanoi specimens. mescaline concentration value of 0.32% obtained in our study for Two apparently closely related cultivars of E. pachanoi were Echinopsis santaensis (Rauh and Backeberg) Friedrich and Rowley examined in this study. One of them, E. pachanoi (Huancabamba (E. santaensis) was virtually identical to that reported recently by long spine), yielded a mescaline concentration of 1.2% in cortical Cjuno et al. (2009). O. Ogunbodede et al. / Journal of Ethnopharmacology 131 (2010) 356–362 361

Echinopsis uyupampensis (Backeberg) Friedrich and Rowley (E. whole, but obtained comparable concentrations that facilitated the uyupampensis) has no published analytical data on its mescaline ranking of 14 species and cultivars of Echinopsis with mescaline content. The same is true of Echinopsis scopulicola (Ritter) Mottram contents that spanned two orders of magnitude, from 0.053% to (E. scopulicola). In this study the mescaline content of E. uyupam- 4.7% by dry weight. The mescaline concentrations reported here pensis by dry weight was found to be 0.053%, the lowest of the largely support the hypothesis that plants with higher mesca- 14 specimens analyzed. It would be reasonable to deduce that an line concentrations are associated with documented shamanic use. extremely large amount of tissue from this taxon would have to The numerous populational, environmental and temporal factors be used to obtain an efficacious psychoactive dose of mescaline. which, either alone or in combination, could affect mescaline levels Thus, it seems unlikely that E. uyupampensis would be used for in Echinopsis species, await future investigation. mescaline-based therapeutic and diagnostic purposes as are other species such as E. pachanoi and E. peruviana. Consistent with this Acknowledgements inference is the fact that we can locate no anthropological reports suggesting those applications for E. uyupampensis. The mescaline The authors gratefully acknowledge the essential help of G. Sul- concentration of 0.82% found for E. scopulicola, on the other hand, livan, L. Wetterauer and M. Gallardo with software applications. M. is in the top third of the results for our samples, placing it in a cluster Kalam made helpful comments on an early draft. We thank Wiley- of high-mescaline-content E. pachanoi taxa. Two other taxa whose VCH Verlag GmbH for permission to use the mass spectrum from mescaline concentrations are reported here for the first time, are (1) Rösner et al. (2007), and the San Diego Museum of Man for permis- Echinopsis pachanoi complex ssp. riomizquensis (T. riomizquensis Rit- sion to use the photograph of the shaman with San Pedro cactus ter), sensu Taylor (2007) (E. pachanoi complex ssp. riomizquensis), (from Sharon, 2000) in the graphical abstract. A grant from the Alvin and (2) Echinopsis pachanoi complex ssp. pallarensis (T. pallarensis A. and Roberta E. Klein Foundation made this work possible. Ritter), sensu Taylor in his treatment of E. pachanoi spp. riomizquensis (Taylor, 2007)(E. pachanoi complex ssp. pallarensis). These two taxa, which, along with other members of the E. pachanoi complex, References are clearly in need of study, show mescaline concentrations of 0.47% and 0.40%, respectively, placing them at the upper end of the lowest Agurell, S., 1969a. Identification of alkaloid intermediates by gas chromatography– one-third of the samples analyzed in the present study. Their use mass spectrometry. I. Potential mescaline precursors in Trichocereus species. Lloydia 32, 40–45. by shamanic practitioners has not been documented. Agurell, S., 1969b. Cactaceae alkaloids. I. Lloydia 32, 206–216. The hypothesis that Echinopsis taxa/cultivars with the highest Agurell, S., Bruhn, J.G., Lundström, J., Svensson, U., 1971. Cactaceae alkaloids. X. tissue concentrations of mescaline would be preferred by indige- Alkaloids of Trichocereus species and some other cacti. Lloydia 34, 183–187. Anon., 2007. Oral statement on current indigenous use of Peruvian E. pachanoi in nous shamanic practitioners, appears to be largely supported by the shamanic practice, by indigenous plant dealer who requested anonymity. results of the present study (Table 2). Of the three plants showing Britton, N.L., Rose, J.N., 1920. The Cactaceae, vol. 2. Dover Publications, New York, the highest percentages of mescaline in dried stem chlorenchyma, 241 pp. Bruhn, J.G., Lundström, J., 1976. A student’s experiment in pharmacognosy: biosyn- the first, E. pachanoi (Matucana), and the third, E. pachanoi (Huan- thesis of mescaline in the cactus Trichocereus pachanoi. American Journal of cabamba short spine), are known to be used by practicing shamans Pharmaceutical Education 40, 159–160. in Peru (Anon., 2007 and Friedberg, 1959, respectively). The plant Cjuno, M., Choquenaira, J., Quispe, P., Serrano, C., Tomaylla, C., 2009. El género Tri- chocereus, ecología y contenido mescalínico. Quepo 23, 38–45. with the second-highest mescaline content, E. pachanoi cv. Juul’s Crosby, D.M., McLaughlin, J.L., 1973. Cactus alkaloids. XIX. Crystallization of mesca- Giant, cannot be meaningfully assessed for indigenous shamanic line HCl and 3-methoxytyramine from Trichocereus pachanoi. Lloydia 36, use because of its lack of collection data, despite its close resem- 416–418. Cruz Sánchez, G., 1948. Estudio farmacológico de la Opuntia cylindrica. Thesis. Insti- blance to material encountered in the Arequipa shamans’ market tuto de Farmacología y Terapéutica, Universidad Nacional Mayor de San Marcos, (Trout, 2005). Thus there are positive reports of shamanic use for E. Lima, Peru, 27 pp. pachanoi from Matucana and Huancabamba, the known employ- Djerassi, C., Liu, L.H., Farkas, E., Lippman, A.E., Lemin, A.J., Geller, L.E., McDonald, R.N., ment of E. peruviana by Peruvian shamans (Reyna Pinedo and Taylor, B.J., 1955. Terpenoids. XI. Investigation of nine cactus species. Isolation of two new triterpenes, stellatogenin and machaeric acid. Journal of the American Flores Garcés, 2001; Carlos Ostolaza, in litt.; a commercial Peru- Chemical Society 55, 1200–1203. vian plant supplier requesting anonymity, in litt.), a noninformative Dobkin de Ríos, M., 1972. Plant hallucinogens and the religion of the Mochica, an status for the E. pachanoi Juul’s Giant cultivar, and a lack of posi- ancient Peruvian people. Economic Botany 31, 189–203. Friedberg, C., 1959. Rapport sommaire sur une mission au Pérou. Journal tive data on shamanic use for the remaining plants that showed d’Agriculture Tropicale et de Botanique Appliquées 6, 439–450. lower mescaline concentrations in this study. That is not to imply Friedrich, H., 1974. Zur Taxonomie und Phylogenie der Echinopsidinae (Trichocere- that some of the plants with lower mescaline content are not inae). IOS Bulletin 3, 79–93. Gennaro, M.C., Gioannini, E., Giacosa, D., Siccardi, D., 1996. Determination of mesca- usable – or even that they are not actually used – indeed, one line in hallucinogenic Cactaceae by ion-interaction HPLC. Analytical Letters 29, Peruvian informant asserted that several species/cultivars of the 2399–2409. genus Echinopsis appear to be viewed and used interchangeably by Gonzalez Huerta, I., 1960. Identificación de la mescalina contenida en el Trichocereus pachanoi (San Pedro). Revista del Viernes Médico [Lima] 11, 133–137. indigenous shamans in the Matucana area, without regard to their Helmlin, H., Brenneisen, R., 1992. Determination of psychotropic phenylalkylamine taxonomic identities (Anon., 2007). But the fact remains that in derivatives in biological matrices by high-performance liquid chromatography the set of Echinopsis cacti with adequate collection data, the Peru- with photodiode-array detection. Journal of Chromatography 593, 87–94. Janot, M.M., Bernier, M., 1933. Essai de localization des alcaloïdes dans le peyotl. vian E. pachanoi cacti with the highest mescaline concentrations Bulletin des Sciences Pharmacologiques 40, 145–153. predominate in current documented shamanic use. Marini-Bettòlo, G.B., Coch Frugoni, J.A., 1958. Determination of mescaline in hallucinogenic Cactaceae. Journal of Chromatography 1, 182–185. Pardanani, J.H., McLaughlin, J.L., Kondrat, R.W., Cooks, R.G., 1977. Alkaloids. XXXVI. Mescaline and related compounds from Trichocereus peruvianus. Lloydia 40, 5. Conclusion 585–590. Poisson, J., 1960. Présence de mescaline dans une Cactacée péruvienne. Annales Echinopsis researchers in the past have examined various tis- Pharmaceutiques Franç aises 18, 764–765. Pummangura, S., McLaughlin, J.L., Schifferdecker, R.C., 1982. Cactus alkaloids. LI. Lack sues of a broad set of taxa/cultivars of plants, employing different of mescaline translocation in grafted Trichocereus. Journal of Natural Products extraction and analytical procedures, all of which may account for 45, 224–225. disparities in alkaloid recoveries. In the present study, by extract- Reti, L., Castrillón, J.A., 1951. Cactus alkaloids. I. Trichocereus terscheckii (Parmentier) Britton and Rose. Journal of the American Chemical Society 73, 1767–1769. ing only the cortical stem chlorenchyma, we almost certainly Reyna Pinedo, V., Flores Garcés, J., 2001. El uso del “San Pedro” (Echinopsis pachanoi) overestimated the average mescaline content of the plant as a en medicina tradicional peruana. Quepo 15, 28–37. 362 O. Ogunbodede et al. / Journal of Ethnopharmacology 131 (2010) 356–362

Rösner, P., Junge, T., Westphal, F., Fritschi, F., 2007. Mass Spectra of Designer Drugs, Shulgin, A., Shulgin, A., 1991. PIHKAL: A Chemical Love Story. Transform Press, Including Drugs, Chemical Warfare Agents, and Precursors, vol. 2. Wiley-VCH Berkeley, CA, 978 pp. Verlag/GmbH & Co./KGaA, Weinheim, 2067 pp. Taylor, N., 2007. Cactaceae of Bolivia: additional department records. Cactaceae Sys- Rowley, G.D., 1974. Reunion of the genus Echinopsis. IOS Bulletin 3, 93–99. tematics Initiatives. Bulletin of the International Cactaceae Systematics Group Schultes, R.E., Hofmann, A., 1980. Botany and Chemistry of Hallucinogens. Charles 22, 10–11. C. Thomas, Springﬁeld, IL, 462 pp. Trout, K., 2005. Trout’s Notes on San Pedro & Related Trichocereus Species, Sacred Serrano, C.A., 2008. Avances en la ﬁtogeografía química del género Trichocereus en Cacti, third ed., Part B. Better Days Publishing, Austin, 310 pp. el sur del Perú. Quepo 22, 29–34. Turner, W.J., Heyman, J., 1960. The presence of mescaline in Opuntia cylindrica. Sharon, D., 2000. Shamanism & the Sacred Cactus. San Diego Museum Papers 37, Journal of Organic Chemistry 25, 2250–2251. San Diego Museum of Man, San Diego, 76 pp. Yetman, D., 2007. The Great Cacti. University of Arizona Press, Tucson, 297 pp.